Abstract 151: Organized 3D Neo-Angiogenic and Neo-Lymphangiogenic Vascular Networks for Cardiac Regeneration

Abstract

Introduction: Cardiovascular tissue engineering has been an area of intense investigation. The major challenge to these approaches has been the inability to vascularize and perfuse the in vitro engineered tissue constructs. Engineering a tissue of clinically relevant magnitude requires the formation of extensive and stable microvascular networks within the tissue. Hypothesis: Functional vascularized cardiac graft can be generated by the interaction of multipotent human mesenchymal stem cells (hMSCs) and human induced pluripotent stem cell-derived embryonic cardiac myocytes (hiPSC-eCMs) in a 3D collagen cell carrier (CCC) scaffold. Methods and Results: To achieve the above aim, we have developed an in vitro 3D functional vascularized cardiac muscle construct using hiPSC-eCMs and hMSCs. Initially, to generate the prevascularized scaffold, human cardiac microvascular endothelial cells (hCMVECs) and hMSCs were co-cultured on CCCs for 7 days under vasculogenic culture conditions. hCMVECs/hMSCs underwent maturation and differentiation characteristic of microvessel morphogenesis and formed extensive plexuses of capillary networks. Next, the hiPSC-eCMs and hMSCs were co-cultured onto these generated prevascularized CCCs for further 7 or 14 days in myogenic culture conditions. hiPSC-eCMS/hMSCs underwent maturation and differentiation, and demonstrated spontaneous rhythmic beating. The vascular and cardiac phenotypic inductions were analyzed at the morphological, immunological, biochemical, molecular and functional levels. Expression analyses of the differentiated cells revealed neo-angiogenesis and/or neo-lymphangiogenesis as well as neo-cardiomyogenesis. Conclusions: Our unique 3D co-culture system provides us the apt in vitro functional prevascularized 3D cardiac graft that can be utilized for myocardial repair and/or regeneration.